Differential morphometric and micro-morpho-anatomical responses toward types of culture vessels used in micropropagation of Hemidesmus indicus (L.) R. Br.

Plantlets of Hemidesmus indicus (L.) R. Br. (Indian sarasaparilla) were developed in vitro in two different types of culture vessels and closure types—(1) culture bottles with polypropylene screw-caps and (2) conical flasks with cotton plugs both containing Murashige and Skoog medium with 1.0 mg L−1 6- benzylaminopurine, 0.5 mg L−1 kinetin, and 0.1 mg L−1 indole-3-acetic acid. Shoots proliferated in the bottles exhibited production of (i) a few leaves with reduced leaf area, (ii) wiry and brittle stem, and (iii) less number of shoots, whereas, the shoots developed in culture flasks were of superior characteristics with healthy leaves. The leaves developed in bottles revealed the presence of under-developed cuticle, lower palisade/spongy ratios, undifferentiated epidermal layers, abnormal and non-functional stomata, absence of periderm, and unorganized ground and vascular tissue systems preceded the development of hyperhydrated shoots. The use of a conical flasks culture system promoted the development of natural anatomical features in plantlets. Overall, the type of culture vessel remarkably influenced the growth and development of in vitro-derived plantlets of H. indicus which could be explored for commercial propagation of this multipotent medicinal plant. Compared with glass bottles, cotton plugged flasks improved shoots formation, morpho anatomical parameters, rooting, and survival of in vitro plantlets of Hemidesmus indicus.